6-[D-α-(Coumarin-3-carboxamido)arylacetamido]-penicillanic acids or salts

ABSTRACT

This disclosure describes compounds of the class of 6-[D-α-(substituted-coumarin-3-carboxamido)phenylacetamido]penicillanic acids which possess antimicrobial activity.

BRIEF SUMMARY OF THE INVENTION

This invention relates to new derivatives of 6-aminopenicillanic acidand, more particularly, is concerned with novel compounds which may berepresented by the following structural formula: ##STR1## wherein R₁ ishydrogen or hydroxy; R₂ and R₄ are each individually selected from thegroup consisting of hydrogen, hydroxy, alkoxy having up to 4 carbonatoms, amino, monoalkylamino having up to 4 carbon atoms anddialkylamino having up to 4 carbon atoms in each alkyl group; R₃ and R₅are each individually selected from the group consisting of hydrogen,hydroxy, alkyl having up to 4 carbon atoms, alkoxy having up to 4 carbonatoms, alkenyl having from 3 to 6 carbon atoms, alkanoyloxy having up to4 carbon atoms, alkoxycarbonyloxy having up to 4 carbon atoms, chloro,bromo, nitro, amino, monoalkylamino having up to 4 carbon atoms,dialkylamino having up to 4 carbon atoms in each alkyl group, carboxy,phenyl, p-chlorophenyl and p-bromophenyl; R₂ and R₃ taken together, R₃and R₄ taken together, and R₄ and R₅ taken together are eachbutadienylene; and M is hydrogen or a pharmaceutically acceptablenon-toxic cation with the proviso that at least two of R₂, R₃, R₄ and R₅must be hydrogen.

DETAILED DESCRIPTION OF THE INVENTION

Suitable alkyl and alkoxy groups contemplated by the present inventionare, for example, methyl, ethyl, isopropyl, sec-butyl, methoxy, ethoxy,n-propoxy, isobutoxy, and the like. Appropriate alkenyl groups may be,for example, allyl, methallyl, isopropenyl, 1-butenyl, crotyl,3-butenyl, etc. Suitable alkanoyloxy and alkoxycarbonyloxy groups are,for example, acetyloxy, propionyloxy, isobutyryloxy, methoxycarbonyloxy,isopropoxycarbonyloxy, isobutoxycarbonyloxy, and the like. Thepharmacologically acceptable cations embraced by M in the above generalformula include, for example, the non-toxic metal cations such as thesodium ion, potassium ion, calcium ion, magensium ion, as well as theorganic amine cations such as the tri(lower alkyl)amine cations (e.g.,triethylamine), procaine, and the like.

The novel compounds of the present invention may be readily prepared bycondensing an ampicillin derivative of the formula: ##STR2## wherein R₁and M are as hereinabove defined with a coumarin-3-carboxylic acidderivative of the formula: ##STR3## wherein R₂, R₃, R₄ and R₅ are ashereinabove defined and X is chloro or bromo as in an acid halide, orthe moiety --O--CO--OC₂ H₅ as in a mixed anhydride from ethylchloroformate, or the moiety --O--C(═N-cyclohexyl)NH-cyclohexyl as in adicyclohexylcarbodiimide mediated reaction, or an azide function, or anazolide formed from carbonyldiimidazole. The acylation of the ampicillinderivative is best performed in an inert solvent such astetrahydrofuran, dioxane, methylene chloride or chloroform (or mixturesthereof) at from ice bath temperature (about 0° C.) to room temperature(about 25° C.). The reaction is preferably carried out in the presenceof an acid acceptor such as N-methylmorpholine, triethylamine, or sodaash and over a period of a few hours or more. The acylating agents maybe prepared by methods well known in the art from the correspondingacids (X is hydroxy). Thus, an acid may be treated with a thionyl halideor oxalyl halide, if desired, in the presence of dimethylformamide, toyield the corresponding acyl halides (X is chloro or bromo), which canbe converted to the acyl azides (X is N₃) by treatment with sodiumazide.

The novel compounds of this invention may be physically characterizedand their purity determined by any or all of the following methods: thinlayer chromatography (t.l.c.), infrared spectroscopy (i.r.), nuclearmagnetic resonance spectroscopy (n.m.r.), and high pressure liquidchromatography (h.p.l.c.). In t.l.c. an 80% n-propanol:20% water systemwith silica gel GF plates is used. Compounds appearing approximately atRf 0.6 are detected by ultraviolet quenching and a toluidine-potassiumiodide spray after chlorine fogging. In addition, these compounds areninhydrin negative, whereas ampicillin [generally at a lower Rf (0.5)]is ninhydrin positive. In the i.r. spectra (potassium bromide disc),these compounds are characterized by peaks at 5.6-5.65μ (β-lactam) 5.8,6 and 6.6μ (lactone and amide carbonyls). In the n.m.r. spectra(D-6-dimethylsulfoxide, 100 MHz), characteristic peaks appear at 9.7δ(d, 1H, amide), 9.3δ (d, 1H, amide), 9.4δ to 8.8δ (S, 1H, 4-H ofcoumarin ring), 8.6δ to 7.2δ (M-aromatic protons of coumarin and phenylrings), 5.9δ (d, 1H, CH of phenylacetyl side chain), 5.55δ (q, 1H, H-6of penam ring), 5.4δ (d, 1H, H-5 of penam ring), 1.55δ (S, 3H, CH₃ ofpenam), 1.4δ (S, 3H, other CH₃ of penam), with additional peakscharacteristic of each compound. The h.p.l.c. is performed using aPartisil ODS (Waters' Assoc.) column with a phosphate buffer (pH7):acetonitrile (85:15) system on a Waters' high pressure instrument. Byone or more of these methods, product purity of the compounds of thisinvention was established at 80-90%.

The novel compounds of the present invention are biologically active andhave been found to possess antibacterial activity. As indicated, theyare useful antimicrobial agents and have broad-spectrum antimicrobialactivity in vitro against standard laboratory microorganisms used toscreen for activity against pathogens. The antibacterial spectrum oftypical compounds of the present invention, representing theconcentration required to inhibit the growth of various typicalbacteria, was determined in a standard manner by the agar-dilutionstreak-plate technique. A Steers multiple inocula replicator wasemployed with incubation at 37° C. for 18 hours in conventional nutrientagar. The results are set forth in Table I below expressed as theminimal inhibitory concentration in micrograms per milliliter.

                                      TABLE I                                     __________________________________________________________________________                    Pseudomonas                                                                          Klebsiella                                                                          Enterobacter                                                                         Proteus                                                                             Proteus                                                                             Escherechia                                                                          Staphylococcus                         aeruginosa                                                                           pneumoniae                                                                          cloacae                                                                              mirabilis                                                                           morganii                                                                            coli   aureus                   Compound      UCS 7613                                                                             MA 75-2                                                                             OSU 75-2                                                                             OSU 75-3                                                                            K 72  CU 75-1                                                                              OSU                    __________________________________________________________________________                                                           75-2                   6-[D-α-(6-Hydroxycoum-                                                  arin-3-carboxamido)-                                                                          2      16    16     4     32    8      0.25                   phenylacetamido]penicil-                                                      lanic acid                                                                    6-[D-α-(7-Methoxycouma-                                                 rin)-3-carboxamido)-                                                                          1      8     16     2     16    8      1                      phenylacetamido]penicil-                                                      lanic acid                                                                    6-[D-α-(8-Nitrocoumarin-                                                3-carboxamido)phenyl-                                                                         8      32    32     1     16    16     1                      acetamido]penicillanic                                                        acid                                                                          6-[D-α-(5-Azacoumarin-                                                  3-carboxamido)phenyl-                                                                         4      128   --     1     128   16     0.5                    acetamido]penicillanic                                                        acid                                                                          6-[D-α-(5,7-Dimethoxy-                                                  coumarin-3-carboxamido)-                                                                      2      128   32     16    64    16     1                      phenylacetamido]peni-                                                         cillanic acid                                                                 6-[D-α-(5,6-Benzocouma-                                                 rin-3-carboxamido)phen-                                                                       2      16    16     2     32    4      0.12                   ylacetamido]penicillanic                                                      acid                                                                          6-[D-α-(8-Allylcoumarin-                                                3-carboxamido)-phenyl-                                                                        2      8     8      2     16    4      0.12                   acetamido]penicillanic                                                        acid                                                                          6-[D-α-(Coumarin-3-car-                                                 boxamido)phenylaceta-                                                                         2      16    16     4     16    8      2                      mido]penicillanic acid                                                        6-[D-α-(5,6-Benzocouma-                                                 rin-3-carboxamido)-p-                                                                         2      64    64     4     64    16     0.5                    hydroxyphenylaceta-                                                           mido] penicillanic acid                                                       6-[D-α-(6-Ethoxycarbo-                                                  nyloxycoumarin-3-carbox-                                                                      2      16    32     4     32    8      0.25                   amido)phenylacetamido]-                                                       penicillanic acid                                                             6-[D-α-(6,8-Dichloro-                                                   coumarin-3-carboxamido)-                                                                      2      8     8      1     8     4      0.12                   phenylacetamido]penicil-                                                      lanic acid                                                                    6-[D-α-(6,8-Dibromo-                                                    coumarin-3-carboxamido)-                                                                      4      8     16     2     8     4      0.25                   phenylacetamido]penicil-                                                      lanic acid                                                                    6-[D-α-(6-Phenylcoumar-                                                 in-3-carboxamido)phen-                                                                        2      4     8      2     8     2      0.12                   ylacetamido]penicillanic                                                      acid                                                                          6-[D-α(6-Chlorocoumar-                                                  in-3-carboxamido)phen-                                                                        2      16    16     2     32    8      0.25                   ylacetamido]penicillanic                                                      acid                                                                          6-[D-α-(6-Carboxycoumar-                                                in-3-carboxamido)phenyl-                                                                      128    >128  >128   64    >128  64     4                      acetamido]penicillanic                                                        acid                                                                          6-[D-α-(6,7-Benzocoumar-                                                in-3-carboxamido)-phen-                                                                       1      8     4      0.5   16    16     ≦0.06           ylacetamido]penicillanic                                                      acid                                                                          6-[D-α-(6-p-chlorophen-                                                 ylcoumarin-3-carboxa-                                                                         16     16    32     8     16    16     1                      mido)phenylacetamido]-                                                        penicillanic acid                                                             6-[D-α-(6-Nitrocoumarin-                                                3-carboxamido)phenyl-                                                                         16     64    128    4     64    32     1                      acetamido]penicillanic                                                        acid                                                                          6-[D-α-(8-Aminocoumarin-                                                3-carboxamido)phenyl-                                                                         32     >128  >128   4     128   64     2                      acetamido]penicillanic                                                        acid                                                                          7-[D-α-(Coumarin-3-car-                                                 boxamido)phenylaceta-                                                                         8      32    32     16    128   32     0.5                    mido)cephalosporanic                                                          acid                                                                          6-[D-α-(7,8-Benzocou-                                                   marin-3-carboxamido)-                                                                         4      16    16     1     16    8      0.5                    phenylacetamido]peni-                                                         cillanic acid                                                                 Carbenicillin   32     >128  32     1     128   8      0.5                    Ampicillin      >128   16    >128   0.25  >128  1      0.12                   6-[D-α-(7,8-Benzocou-                                                   marin-3-carboxamido-                                                                          2      16    8      4     16    8      0.12                   phenylacetamido]peni-                                                         cillanic acid                                                                 6-[D-α-(7-Diethyla-                                                     minocoumarin-3-carbox-                                                                        1      4     4      4     8     8      0.25                   amido)phenylacetami-                                                          do]penicillanic acid                                                          7-[D-α-(Coumarin-3-                                                     carboxamido)phenyl-                                                                           4      16    16     4     32    32     0.5                    acetamido]-3-[(1-                                                             methyltetrazole-5-                                                            thio)methyl]ceph-3-                                                           em-4-carboxylic                                                               acid                                                                          __________________________________________________________________________

A preferred embodiment of the present invention may be represented bythe following structural formula: ##STR4## wherein R₁ and R₃ are eachindividually selected from the group consisting of hydrogen, hydroxy,methoxy and ethoxy; R₂ and R₄ are each individually selected from thegroup consisting of hydrogen, hydroxy, methyl, ethyl, methoxy, ethoxy,allyl, methallyl, nitro, amino, chloro, bromo and phenyl; R₁ and R₂taken together, R₂ and R₃ taken together; and R₃ and R₄ taken togetherare each butadienylene; and M is hydrogen or a pharmacologicallyacceptable cation with the proviso that at least two of R₁, R₂, R₃ andR₄ must be hydrogen.

A most preferred embodiment of the present invention may be representedby the following structural formula: ##STR5## wherein R' and R" are eachindividually selected from the group consisting of hydrogen, hydroxy,methoxy, ethoxy, chloro and bromo and M is hydrogen or apharmaceutically acceptable cation.

The invention will be described in conjunction with the followingspecific examples.

EXAMPLE 16-[D-α-(5,6-Benzocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

To a suspension of 483 mg. of 5,6-benzocoumarin-3-carboxylic acid in 45ml. of dioxane and 10 ml. of acetone is added 0.282 ml. oftriethylamine. The mixture is cooled and stirred in an ice bath and0.194 ml. of ethyl chloroformate is added dropwise over 10 minutes.Stirring and cooling is continued another 45 minutes. Then 0.281 ml. oftriethyl amine and 807 mg. of ampicillin trihydrate are added. Stirringand cooling is continued another hour. A chilled mixture of 3 ml. ofsaturated sodium bicarbonate solution and 35 ml. of water is added andthe reaction mixture is shaken with 100 ml. of ethyl acetate. Themixture is filtered to remove suspended solids. The aqueous layer isacidified, with cooling, with 6 N hydrochloric acid to pH 2 and thenextracted with 100 ml., then 50 ml. of ethyl acetate. The combined ethylacetate extracts are washed with 25 ml. of water, dried over magnesiumsulfate and evaporated to a solid. Trituration with ether and filtrationgives the desired product, i.r. 5.4μ (β lactam).

EXAMPLE 26-[D-α-(8-Allylcoumarin-3-carboxamido)phenylacetamido]penicillanic acid

To a stirred solution of 430 mg. of 8-allylcoumarin-3-carboxylic acid[R. R. Bui-Hoi, et al., Bull. Soc. Chim. Fr., 128 (1947)] and 0.225 ml.of N-methylmorpholine in 25 ml. of methylene chloride and 25 ml. ofdioxane, cooled in an ice-methanol bath to -10° to -12° C., is added0.192 ml. of ethyl chloroformate dropwise. After 8 minutes, 0.223 ml. ofN-methylmorpholine and 807 mg. of ampicillin trihydrate are added. Theice-methanol bath is replaced with an ice bath, and after one additionalhour a chilled mixture of 20 ml. of dioxane, 10 ml. of acetone, 35 ml.of water and 3 ml. of saturated sodium bicarbonate is added. Thereaction mixture is worked up as described in Example 1 (except nofiltration of the two phase mixture is necessary). Trituration of theproduct with ether-hexane and filtration gives the desired product, i.r.5.62μ (β lactam).

EXAMPLE 3 6-[D-α-(Coumarin-3-carboxamido)phenylacetamido]-penicillanicacid

This product is prepared as described in Example 2, except that thestarting material is 382 mg. of coumarin-3-carboxylic acid. The desiredproduct is obtained, i.r. 5.67μ (β lactam).

EXAMPLE 46-[D-α-(5,6-Benzocoumarin-3-carboxamido)-p-hydroxyphenylacetamido]penicillanicacid

To a suspension of 483 mg. of 5,6-benzocoumarin-3-carboxylic acid and0.225 ml. of N-methylmorpholine in 25 ml. of dioxane and 25 ml. ofmethylene chloride, cooled to -10° to -15° C. in an ice-methanol bath,is added 0.192 ml. of ethyl chloroformate dropwise over several minutes.After an additional 25 minutes of stirring and cooling, 0.223 ml. ofN-methylmorpholine and 770 mg. of amoxicillin are added. The mixture isstirred in an ordinary ice bath for one hour. Then a cold mixture of 20ml. of dioxane, 10 ml. of acetone, 35 ml. of water and 3 ml. ofsaturated sodium bicarbonate is added. Workup is done as described inExample 1 giving the desired product, i. 4. 5.65μ (β lactam).

EXAMPLE 56-[D-α-(6-Ethoxycarbonyloxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid

This product is prepared as described in Example 2, except that 417 mg.of 6-hydroxycoumarin-3-carboxylic acid [F. D. Cramer & H. Windel, Chem.Ber., 89, 354 (1956)], 0.564 ml. of triethylamine, 0.388 ml. of ethylchloroformate, ice bath (0°-5° C.) temperature and a one hour reactiontime are used to form the mixed anhydride. Reaction with ampicillin andworkup gives the desired product, i.r. 5.67μ (β lactam).

EXAMPLE 66-[D-α-(6-Hydroxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid

This product is prepared as described in Example 5, except that 0.225ml. of N-methylmorpholine, 0.192 ml. of ethyl chloroformate, and areaction time of 15 minutes at -10° C. plus 30 minutes at 0° C. are usedto form the mixed anhydride. Reaction with ampicillin (using 0.223 ml.of N-methylmorpholine) and workup as in Example 5 gives 343 mg. of crudeproduct. Removal of some starting 6-hydroxycoumarin-3-carboxylic acidfrom the product is accomplished by stirring 243 mg. of crude productwith ethyl acetate, filtering and evaporating the ethyl acetate todryness giving the purified product, i.r. 5.66μ (β lactam).

EXAMPLE 76-[D-α-(7-Methoxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid

To a solution of 444 mg. of 7-methoxycoumarin-3-carboxylic acid [W.Baker & C. B. Collis, J. Chem. Soc., 5, 12-15 (1949)] and 0.282 ml. oftriethylamine in 25 ml. of dioxane and 25 ml. of methylene chloride,stirred and cooled in an ice bath, is added 0.194 ml. of ethylchloroformate dropwise. After 25 minutes, 0.281 ml. of triethylamine and807 mg. of ampicillin trihydrate are added. After an additional one hourof reaction at 0°-5° C., workup as described in Example 4 gives thedesired product, i.r. 5.61μ (β-lactam).

EXAMPLE 86-[D-α-(8-Nitrocoumarin-3-carboxamido)phenylacetamido]-penicillanic acid

This reaction is performed as described in Example 7, using 475 mg. of8-nitrocoumarin-3-carboxylic acid [R. O. Clinton and S. G. Laskowski, J.Am. Chem. Soc., 71, 3662 (1949)]. The desired product is obtained, i.r.5.60μ (β-lactam).

EXAMPLE 96-[D-α-(5-Azacoumarin-3-carboxamido)phenylacetamido]penicillanic acid

This reaction is performed as described in Example 7, using 386 mg. of5-azacoumarin-3-carboxylic acid [R. B. Moffitt, J. Org. Chem., 35, 3596(1970)]. The desired product is obtained, i.r. 5.63μ (β-lactam).

EXAMPLE 106-[D-α-(5,7-Dimethoxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid

A 0.5 g. portion of 5,7-dimethoxycoumarin-3-carboxylic acid [A.Roberston and T. S. Subramaniam, J. Chem. Soc., 286 (1937)] is reactedwith 0.195 ml. of ethyl chloroformate and 0.28 ml. of triethylamine.Then 0.28 ml. of triethylamine ethylamine and 0.8 g. of ampicillintrihydrate are added according to the procedure of Example 1 giving thedesired product, i.4. 5.61μ (β lactam).

EXAMPLE 116-[D-α-(6,8-Dichlorocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

A 0.518 g. portion of 6,8-dichlorocoumarin-3-carboxylic acid [Ng. Ph.Bui-Hoi, et al., Bull. Soc. Chim. Fr., 561 (1957)] is reacted with 0.195ml. of ethyl chloroformate and 0.28 ml. of triethylamine. Then 0.28 ml.of triethylamine and 0.8 g. of ampicillin trihydrate are added accordingto the procedure of Example 1, giving the desired product, i.r. 5.65μ (βlactam).

EXAMPLE 126-[D-α-(6,8-Dibromocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

A 0.692 g. portion of 6,8-dibromocoumarin-3-carboxylic acid [Ng. Ph.Bui-Hoi, et al., Bull. Soc. Chim. Fr., 561 (1957)] is reacted with 0.195ml. of ethylchloroformate and 0.28 ml. of triethylamine. Then 0.28 ml.of triethylamine and 0.8 g. of ampicillin trihydrate are added accordingto the procedure of Example 1, giving the desired product, i.r. 5.63μ (βlactam).

EXAMPLE 136-[D-α-(6-Phenylcoumarin-3-carboxamido)phenylacetamido]penicillanic acid

A 0.27 g. portion of 6-phenylcoumarin-3-carboxylic acid is reacted with0.095 ml. of ethyl chloroformate and 0.14 ml. of triethylamine. Then0.14 ml. of triethylamine and 0.4 g. of ampicillin trihydrate are addedaccording to the procedure of Example 1, giving the desired product,i.r. 5.67μ (β lactam).

EXAMPLE 146-[D-α-(6-Chlorocoumarin-3-carboxamido)phenylacetamido]penicillanic acid

A 0.45 g. portion of 6-chlorocoumarin-3-carboxylic acid [L. L. Woods andJ. Sapp, J. Org. Chem., 30, 312 (1965)] is reacted with 0.195 ml. ofethyl chloroformate and 0.28 ml. of triethylamine. Then 0.28 ml. oftriethylamine and 0.8 g. of ampicillin trihydrate are added according tothe procedure of Example 1, giving the desired product, i.r. 5.68μ (βlactam).

EXAMPLE 156-[D-α-(6-Carboxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid

To a suspension of 400 mg. of 6-carboxycoumarin-3-carboxylic acid [R.Selleri, Arzn. Forsch., 15, 910 (1965)] and 0.38 ml. ofN-methylmorpholine in 25 ml. of dioxane and 25 ml. of methylenechloride,cooled to -15° C. in an ice-methanol bath, is added 0.323 ml. of ethylchloroformate dropwise. Then more solvent is added and the reaction heldat 0° C. for 30 minutes. Addition of 0.19 ml. of N-methylmorpholine and685 mg. of ampicillin trihydrate gives, after the two isomeric compoundsderived from reaction of either the 6- or 3-carboxy group of thestarting acid. Separation by chromatography gives the desired product,i.r. 5.62μ (β lactam).

EXAMPLE 166-[D-α-(6,7-Benzocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

This compound is prepared similarly to the 5.6-benzo analog ofExample 1. Thus, 336 mg. of carboxylic acid [T. Boehm and E. Profft,Arch. Pharm., 269, 25 (1931)] gives the desired compound, i.r. 5.67μ (βlactam).

EXAMPLE 176-[D-α-(6-p-Chlorophenylcoumarin-3-carboxamido)phenylacetamido]penicillanicacid

This compound is prepared similarly to Example 1. Thus, 202 mg. ofcarboxylic acid gives the desired compound, i.r. 5.63μ (β lactam).

EXAMPLE 18 6-(p-Chlorophenyl)coumarin-3-carboxylic acid

4-(p-Chlorophenyl)phenol is converted via the Duff reaction [J. Chem.Soc., 547 (1941)] to 2-hydroxy-5-(p-chlorphenyl)benzaldehyde, mp.74°-78° C. i.r. 6.02μ. A solution of 233 mg. (1 mmole) of this aldehyde,0.4 ml. (2.63 mmole) of diethylmalonate and 0.2 ml. (2 mmole) ofpiperidine is heated for 10 minutes and kept at 25° C. for 2 hours. Then5 ml. of 9 N HCl is added and the mixture stirred at reflux overnight.The mixture is cooled in an ice bath and filtered to yield 238 mg. ofthe desired compound, i.r. 5.74, 5.9μ.

EXAMPLE 196-[D-α-(6-Nitrocoumarin-3-carboxyamido)phenylacetamido]penicillanic acid

This compound is prepared similarly to the 8-nitro analog of Example 8to give the desired product, i.r. 5.60μ (β lactam). The6-nitrocoumarin-3-carboxylic acid is obtained after Lampe andMacierwicz, Roezniki Chem., 18, 668 (1938).

EXAMPLE 206-[D-α-(8-Aminocoumarin-3-carboxamido)phenylacetamido]penicillanic acid

This compound is prepared by reduction of 125 mg. of the 8-nitro analogof Example 8 with hydrogen and 25 mg. of 10% Pd/C in ethanol to give thedesired product, i.r. 5.65μ (β lactam).

EXAMPLE 217-[D-α-(Coumarin-3-carboxamido)phenylacetamido]cephalosporanic acid

To a solution of 190 mg. of coumarin-3-carboxylic acid and 0.141 ml. oftriethylamine in 12 ml. of dioxane and 12 ml. of methylene chloride isadded 0.097 ml. of ethyl chloroformate at 0°-5° C. After 30 minutes,0.141 ml. of triethylamine and one mmole of cephaloglycin are added.After one hour at 5° C., typical workup gives the desired product, i.r.5.64μ (β lactam).

EXAMPLE 226-[D-α-(7,8-Benzocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

This compound is prepared as described in Example 1, i.r. 5.65μ (βlactam). The acid is prepared after F. D. Cramer and H. Windel, Chem.Ber., 89, 354 (1956).

EXAMPLE 236-[D-α-(6-Methylcoumarin-3-carboxamido)phenylacetamido]penicillanic acid

From 412 mg. of 6-methylcoumarin-3-carboxylic acid [B. Reichert and W.Hoss, Arch. Pharm., 280, 157 (1942)], 0.282 ml. of triethylamine and0.194 ml. of ethylchloroformate is prepared the mixed anhydride.Reaction with 0.281 ml. of triethylamine and 807 mg. of ampicillin.3H₂ Ogives 567 mg. of product.

EXAMPLE 246-[D-α-(7-Diethylaminocoumarin-3-carboxamido)phenylacetamido]penicillanicacid

To a solution of 527 mg. (2.02 mmoles) of7-diethylaminocoumarin-3-carboxylic acid [mp. 227°-229° C., fromhydrolysis of ethyl 7-diethylaminocoumarin-3-carboxylate (J. D. Kendall,et al., Ilford Ltd., Brit. Pat. 867,592; Chem. Abstr., 55, 21927-C)],0.282 ml. (2.04 mmoles) of triethylamine in 25 ml. of methylene chlorideand 25 ml. of dioxane is added 0.194 ml. (2.04 mmole) ofethylchloroformate, dropwise, at 0°-5° C. over 10 minutes. After 30minutes, 0.281 ml. of triethylamine and 807 mg. of ampicillin trihydrateare added and the mixture stirred for one hour. The aqueous phase froman aqueous bicarbonate-ethyl acetate quench of the reaction mixture isacidified to pH 2.5. Extraction with ethyl acetate and evaporation gives773 mg. of the desired product.

EXAMPLE 257-[D-α-(Coumarin-3-carboxamido)phenylacetamido]-3-[(1-methyltetrazole-5-thio)methyl]ceph-3-em-4-carboxylicacid

A mixture of 288 mg. (0.5 mmole) of the compound described in Example21, 84 mg. (1 mmole) of sodium bicarbonate and 87 mg. (0.75 mmole) of1-methyltetrazole-5-thiol in 8 ml. of 0.1 M pH 6.4 phosphate buffer and10 ml. of acetone is heated under reflux at 50°-55° C. (oil bath) for 3hours. Then an additional 58 mg. of the tetrazole and 60 mg of sodiumbicarbonate (in 5 drops of water) are added and the solution heatedovernight. The acetone is removed in vacuo and the cooled aqueousresidue acidified to pH 2.2 to precipitate a white solid. This isfiltered off, washed with water and dried to give 262 mg., i.r. (KBr)5.60μ (β lactam).

We claim:
 1. A compound selected from the group consisting of6-[D-α-(8-methoxycarbonyloxycoumarin-3-carboxamido)phenylacetamido]penicillanicacid and the pharmacologically acceptable cationic salts thereof.
 2. Acompound selected from the group consisting of6-[D-α-(5-azacoumarin-3-carboxamido)phenylacetamido]penicillanic acidand the pharmacologically acceptable cationic salts thereof.